LOCUS       sequence-345290-       10428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting vector for generating Sox2 STREAMING-tag KI cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10428)
  AUTHORS   Ohishi H, Shimada S, Uchino S, Li J, Sato Y, Shintani M, Owada H, 
            Ohkawa Y, Pertsinidis A, Yamamoto T, Kimura H, Ochiai H
  TITLE     STREAMING-tag system reveals spatiotemporal relationships between 
            transcriptional regulatory factors and transcriptional activity.
  JOURNAL   Nat Commun. 2022 Dec 20;13(1):7672. doi: 10.1038/s41467-022-35286-2.
  PUBMED    36539402
REFERENCE   2  (bases 1 to 10428)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2022-12-20"; volume: "13(1):7672. doi"; pages: " 
            10.1038/s41467-022-35286-2"
COMMENT     Sequence Label: pTV-Sox2_5prime-1000-24MS96T-3F_NeoR
FEATURES             Location/Qualifiers
     source          1..10428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2164..2173
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2476..2494
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2505..2523
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2534..2552
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2579..2597
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2608..2626
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2637..2655
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2682..2700
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2711..2729
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2740..2758
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2785..2803
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2814..2832
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2843..2861
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2888..2906
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2917..2935
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2946..2964
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2991..3009
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3020..3038
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3049..3067
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3094..3112
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3123..3141
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3152..3170
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3197..3215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3226..3244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3255..3273
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3300..3318
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3329..3347
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3358..3376
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3403..3421
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3432..3450
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3461..3479
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3506..3524
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3535..3553
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3564..3582
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3609..3627
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3638..3656
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3667..3685
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3712..3730
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3741..3759
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3770..3788
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3815..3833
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3844..3862
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3873..3891
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3918..3936
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3947..3965
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3976..3994
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4021..4039
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4050..4068
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4079..4097
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4124..4142
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4153..4171
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4182..4200
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4227..4245
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4256..4274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4285..4303
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4330..4348
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4359..4377
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4388..4406
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4433..4451
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4462..4480
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4491..4509
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4536..4554
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4565..4583
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4594..4612
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4639..4657
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4668..4686
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4697..4715
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4742..4760
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4771..4789
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4800..4818
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4845..4863
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4874..4892
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4903..4921
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4948..4966
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4977..4995
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5006..5024
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5051..5069
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5080..5098
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5109..5127
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5154..5172
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5183..5201
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5212..5230
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5241..5259
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5286..5304
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5315..5333
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5344..5362
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5389..5407
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5418..5436
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5447..5465
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5492..5510
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5521..5539
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5550..5568
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5595..5613
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5624..5642
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5653..5671
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    5738..5763
                     /label=SA
                     /note="splice acceptor site"
     CDS             6129..6152
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6168..6191
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6207..6230
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(8248..8268)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8248..8266)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8287..8303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8287..8303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8300..8322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8311..8327
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8335..8341,8342..8359,8360..8365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8380..8401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8511..8528)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8682..9270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8762..8781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9441..10232,10233..10301))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     10064..10083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10302..10392)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggtttgaca
      661 tgcctgcaag tgcagggagc gcagtgccgc ggatgagcgc agaaacaatg gcacaccacc
      721 tccggctctg ccagcttcct gaaatactag ttggacagtc gccctgaacc acccatgggc
      781 cttgccccac cctggcccca gcttcccgcg ccccatccac ccttatgtat ccaagagaga
      841 gccaatattc cgtagcatgg gggaaaggag ctgtcgtctt ggtgctgttt acccacttcc
      901 ttcgaacagg cgtgcgccgt gacctgttgc tgaaaacggg gggcgggggg gggggataca
      961 aaggttcccc agcggccggc tgcgggcccg cctcccccgc gcggttcggg gcacagcgct
     1021 ctgctgggct cggctcggcg gcgcggcagg ccccgccccc tttcatgcaa aaccctctgg
     1081 cgaggctggg ctcgggcgca ggagccggcg ctcgctgatt ggccgccgga aacccattta
     1141 ttccctgaca gcccccatca catggatggt tgtctattaa cttgttcaaa aaagtatcag
     1201 gagttgtcaa ggcagagaag agagtgtttg caaaaaggga aaagtacttt gctgcctctt
     1261 taagactagg gctgggagaa agaagaggag agagaaagaa aggagagaag tttggagccc
     1321 gaggcttaag cctttccaaa aactaatcac aacaatcgcg gcggcccgag gaggagagcg
     1381 cctgtttttt catcccaatt gcacttcgcc cgtctcgagc tccgcttccc cccaactatt
     1441 ctccgccaga tctccgcgca gggccgtgca cgccgaggcc cccgcccgcg gcccctgcat
     1501 cccggccccc gagcgcggcc cccacagtcc cggccgggcc gagggttggc ggccgccggc
     1561 gggccgcgcc cgcgcccagc gcccgcatgt ataacatgat ggagacggag ctgaagccgc
     1621 cgggcccgca gcaagcttcg gggggcggcg gcggaggagg caacgccacg gcggcggcga
     1681 ccggccaggt aagcgataag tattcgatca tgcattggtt agcggccgcg aattcgccct
     1741 tggatccgga atgaccacca ggcattccga tgccacgatg gaccatcagg ccatcgagct
     1801 gcagaagtga cgaccacgca cttcggagtg acagaggagg atcacccctc tggccaccag
     1861 agtagagcat cagcctactc acagagccta cggaggacca ccccgtagga gtagagcgag
     1921 gagcaccagc cctcgcgtgt gacgcacgag catcagccgt gcgatggtaa gatccggaat
     1981 gaccaccagg cattcctcca aggcgatgga ccatcaggcc atcgaggact cgaagtgacg
     2041 accacgcact tcgtgaaggc agaggaggat cacccctctg cccacaggag tagagcatca
     2101 gcctactcca agcagctacg gaggaccacc ccgtagaagt agggcgagga gcaccagccc
     2161 tcgccatggt ggcacgagca tcagccgtgc agggtataga tccggaatga ccaccaggca
     2221 ttcctggcca acgatggacc atcaggccat cggcatagcg aagtgacgac cacgcacttc
     2281 agggtagcag aggaggatca cccctctggc cacacgagta gagcatcagc ctactccaca
     2341 ggactacgga ggaccacccc gtagaaagtg ggcgaggagc accagccctc gccgtgatgg
     2401 cacgagcatc agccgtgcgt taggaagatc tcgattcgat gcagcctagt taggggtcga
     2461 ctagtctact cactatctct atcactgata gggaaacctt cctctctcta tcactgatag
     2521 ggaccttttc ccttctctat cactgatagg gacccatatt ctcctagttt aacttctatc
     2581 tctatcactg atagggaaac tcacttctct ctatcactga tagggacctt atcccctctc
     2641 tatcactgat agggatatta tatttcctag tattcacctc ctctctatca ctgataggga
     2701 ttcaccattt tctctatcac tgatagggat atttccttct ctctatcact gatagggaca
     2761 tatttttccc tagtcaatcc tcactctcta tcactgatag ggactaacaa aattctctat
     2821 cactgatagg gaaaaccaaa catctctatc actgataggg atcatcattt tcctagtata
     2881 accatactct ctatcactga tagggatcac ccccactctc tatcactgat agggattaca
     2941 caacttctct atcactgata gggactctct ttatcctagt cacattaaca tctctatcac
     3001 tgatagggac accttttcat ctctatcact gatagggaca tccattattc tctatcactg
     3061 atagggacat taactctcct agtaaaatcc ccatctctat cactgatagg gaaacctcat
     3121 catctctatc actgataggg aatctattcc atctctatca ctgataggga tatcttttat
     3181 cctagtccta acatcatctc tatcactgat agggaaccca aactttctct atcactgata
     3241 gggaccaatt tacctctcta tcactgatag ggaatcttct tatcctagta caaaactaat
     3301 ctctatcact gatagggaat ccctcccttc tctatcactg atagggatac ctttccttct
     3361 ctatcactga tagggaatta cttttaccta gtaatacctc aatctctatc actgataggg
     3421 aaaatcacta atctctatca ctgataggga tttttattat tctctatcac tgatagggaa
     3481 ctcctttaac ctagtctatt ccatttctct atcactgata gggattcact accctctcta
     3541 tcactgatag ggattatttc ttctctctat cactgatagg gaatttttcc ttcctagtac
     3601 cataaccctc tctatcactg atagggaaca ccaatattct ctatcactga tagggacatt
     3661 atttattctc tatcactgat agggaattat tactccctag ttcttacatt atctctatca
     3721 ctgataggga acccctctcc tctctatcac tgatagggat atcaatcact ctctatcact
     3781 gatagggacc tattataccc tagtccaata tatatctcta tcactgatag ggaccacact
     3841 tcttctctat cactgatagg gatcacctta tatctctatc actgataggg acactctcta
     3901 tcctagtcca atccttttct ctatcactga tagggataaa cacatctctc tatcactgat
     3961 agggatacat tttactctct atcactgata gggattctcc ctttcctagt cttaacttct
     4021 tctctatcac tgatagggat ttttccaatt ctctatcact gatagggaat cacttcattc
     4081 tctatcactg atagggatta tttctcccct agttcactcc tactctctat cactgatagg
     4141 gactcccaat attctctatc actgataggg atatttttct ctctctatca ctgataggga
     4201 cacctcctct cctagttacc acaacttctc tatcactgat agggaatact ctcaatctct
     4261 atcactgata gggatttcta tttctctcta tcactgatag ggaataccct ctacctagta
     4321 acaaatcttt ctctatcact gatagggaat aatatatctc tctatcactg atagggatct
     4381 tataccctct ctatcactga tagggacctt ccacctccta gttaatcaaa tatctctatc
     4441 actgataggg ataatctact ttctctatca ctgataggga aactcatcca tctctatcac
     4501 tgatagggat ttttaacttc ctagtatcca ttctatctct atcactgata gggataatac
     4561 aaactctcta tcactgatag ggatactctc ctttctctat cactgatagg gaccttattt
     4621 tccctagtat aatccctatc tctatcactg atagggaaca ttaactatct ctatcactga
     4681 tagggatttt attccttctc tatcactgat agggaactta cctatcctag tatacccata
     4741 atctctatca ctgataggga aaccaaaatt tctctatcac tgatagggaa tccaattact
     4801 ctctatcact gatagggatt tctatctccc tagtttaaaa ccaatctcta tcactgatag
     4861 ggaaccccta aactctctat cactgatagg gatttctctt attctctatc actgataggg
     4921 accttctctc acctagtcta ctcactatct ctatcactga tagggaaacc ttcctctctc
     4981 tatcactgat agggaccttt tcccttctct atcactgata gggacccata ttctcctagt
     5041 acactataaa tctctatcac tgatagggaa aacatacctt ctctatcact gatagggact
     5101 atattttttc tctatcactg atagggatct aatacttcct agtactttaa atatctctat
     5161 cactgatagg gaatctacta aatctctatc actgataggg acccataaac ttctctatca
     5221 ctgataggga atttttttcc tctctatcac tgatagggat tatctcttcc ctagttcctc
     5281 ttaaatctct atcactgata gggatctcac tacttctcta tcactgatag ggaaactcat
     5341 aactctctat cactgatagg gaaaattatc ttcctagtaa aacacctatc tctatcactg
     5401 atagggatat tataccatct ctatcactga tagggattta tcattttctc tatcactgat
     5461 agggattttt accatcctag ttaaaaatat ctctctatca ctgataggga tcactaactc
     5521 tctctatcac tgatagggaa ctattctatt ctctatcact gatagggaca ctttttaacc
     5581 tagtacatta cccatctcta tcactgatag ggaacctatc tcttctctat cactgatagg
     5641 gatatttttt catctctatc actgataggg aaaactttac acctaggatt accctgttat
     5701 ccctactcga catcgattag ctgatacgat aaagcttctg acctcttctc ttcctcccac
     5761 agggatgtct gagctacgat acagagatcc tcacagttga gtacggattt ctgcccattg
     5821 gcaaaatagt cgaggaaagg attgaatgca cagtttacac agtggacaag aatggctttg
     5881 tctatacaca gccaatagcc cagtggcata accgaggaga gcaagaggta ttcgagtatt
     5941 gtctcgaaga tgggagtatc atccgagcca ccaaagacca caaattcatg acaacagacg
     6001 gacaaatgct gcctattgac gaaatatttg agagaggttt ggatctgaag caggttgacg
     6061 ggctgcccgc cggagccggt gccggagctg caagcgggaa agagaaaaca catgtttccc
     6121 ggcctaggga ttacaaggat gacgacgata agggcactgc aagcagtgac tataaggatg
     6181 atgatgacaa agggtctggt gggggcgact ataaggatga cgatgacaag cggcctaggc
     6241 ttaatagcaa catggacgca gacctttatg gatataagtg ggctcgcgac aatgtaggac
     6301 agagcggcgc tactatttat cggctctatg gaaaacccga tgctcccgag ctgttcctga
     6361 agcacggcaa gggatccgta gcaaacgacg ttacagacga aatggtacga ctcaactggt
     6421 tgacagaatt tatgcccctg cccactatca agcacttcat aagaactcct gatgacgctt
     6481 ggctgctgac aaccgcaatc cccggtaaaa cagcattcca ggtccttgaa gagtatccag
     6541 actctggtga gaatattgtg gacgcactgg cagtattcct caggcggctc cacagtattc
     6601 ctgtttgcaa ttgtcccttt aactctgaca gggtattcag gctcgcccag gcccaatcac
     6661 gaatgaataa tggcttggtt gatgcaagcg atttcgatga tgaacgcaat ggctggcctg
     6721 ttgaacaagt ttggaaagaa atgcacaagc tcctgccctt ttcacccgat tctgtagtga
     6781 ctcacgggga cttcagcctc gacaatttga tcttcgatga aggaaagctt atcggctgca
     6841 tagatgttgg gcgcgttggg atagccgaca gataccagga cctggccata ttgtggaatt
     6901 gtctcggcga gttttcccct agcttgcaaa aacggctctt tcagaagtat ggcatagata
     6961 accccgacat gaataaactt caattccatc tgatgcttga cgagttcttc ggaactgctg
     7021 gagctggagc tggggctgta aagggaggag gtggatctgg tggaggaggt agtggaggag
     7081 gtggaagtgg tgggggagta aagatcatca gtcgaaaatc cctgggcacc caaaatgtat
     7141 acgatattgg ggtcgagaaa gatcacaatt ttctcctgaa gaatggtctg gttgcaagca
     7201 attgcttcgg caaccagaag aacagcccgg accgcgtcaa gaggcccatg aacgccttca
     7261 tggtatggtc ccgggggcag cggcgtaaga tggcccagga gaaccccaag atgcacaact
     7321 cggagatcag caagcgcctg ggcgcggagt ggaaactttt gtccgagacc gagaagcggc
     7381 cgttcatcga cgaggccaag cggctgcgcg ctctgcacat gaaggagcac ccggattata
     7441 aataccggcc gcggcggaaa accaagacgc tcatgaagaa ggataagtac acgcttcccg
     7501 gaggcttgct ggcccccggc gggaacagca tggcgagcgg ggttggggtg ggcgccggcc
     7561 tgggtgcggg cgtgaaccag cgcatggaca gctacgcgca catgaacggc tggagcaacg
     7621 gcagctacag catgatgcag gagcagctgg gctacccgca gcacccgggc ctcaacgctc
     7681 acggcgcggc acagatgcaa ccgatgcacc gctacgacgt cagcgccctg cagtacaact
     7741 ccatgaccag ctcgcagacc tacatgaacg gctcgcccac ctacagcatg tcctactcgc
     7801 agcagggcac ccccggtatg gcgctgggct ccatgggctc tgtggtcaag tccgaggcca
     7861 gctccagccc ccccgtggtt acctcttcct cccactccag ggcgccctgc caggccgggg
     7921 acctccggga catgatcagc atgtacctcc ccggcgccga ggtgccggag cccgctgcgc
     7981 ccagtagact gcacatggcc cagcactacc agagcggccc ggtgcccggc acggccatta
     8041 acggcacact gcccctgtcg cacatgtgag ggctggactg cgaactggag aaggggagag
     8101 attttcaaag agatacaagg gaattgggag gggtgcaaaa agaggagagt aggaaaaatc
     8161 tgataatgct caaaaggaaa aaaaatctcc gcagcgaaac gacagctgcg gcctgcactt
     8221 gcaggcatgt caaaccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     8281 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     8341 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     8401 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     8461 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggccgcttc ctcgctcact
     8521 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     8581 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     8641 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     8701 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     8761 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     8821 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     8881 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     8941 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     9001 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     9061 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     9121 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     9181 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     9241 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     9301 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     9361 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     9421 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     9481 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     9541 gagggcttac catctggccc cagtgctgca atgataccgc ggctgccacg ctcaccggct
     9601 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     9661 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     9721 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     9781 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     9841 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     9901 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     9961 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
    10021 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
    10081 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
    10141 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
    10201 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
    10261 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct tttcaatatt
    10321 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
    10381 aaaataaaca aatagggatt ccgcgcacat ttccccgaaa agtgccac
//
